Pilot study to evaluate isolation by size of circulating tumour cells in canine oral melanoma.

Liquid biopsy for circulating tumour cell (CTC) detection is generally unexplored in veterinary medicine. Dogs with highly aggressive and heterogeneous tumours, such as oral malignant melanoma (OMM), could benefit from studies involving size-based isolation methods for CTCs, as they do not depend on specific antibodies. This pilot study aimed to detect CTCs from canine OMM using Isolation by Size of Epithelial Tumor Cells (ISET), a microfiltration methodology, followed by immunocytochemistry (ICC) with Melan-A, PNL2, and S100 antibodies. Ten canine patients diagnosed by histopathology and confirmed as OMM by immunohistochemistry were enrolled, their prognostic data was assessed, and blood samples were collected for CTC analysis. Results have shown the detection of intact cells in 9/10 patients. ICC has shown 3/9 Melan-A-positive, 3/9 PNL2-positive, and 8/9 S100-positive patients, confirming the importance of opting for a multimarker assay. A significant number of negative-stained CTCs were found, suggesting their high heterogeneity in circulation. Microemboli stained with either PNL2 or S100 were found in a patient with a high isolated cell count and advanced clinical stage. Preliminary statistical analysis shows a significant difference in CTC count between patients with and without lymph node metastasis (p < .05), which may correlate with tumour metastatic potential. However, we recommend further studies with more extensive sampling to confirm this result. This pilot study is the first report of intact CTC detection in canine OMM and the first application of ISET in veterinary medicine, opening new possibilities for liquid biopsy studies in canine OMM and other tumours.

Tissue and serum microRNA profile of oral squamous cell carcinoma patients.

Head and neck cancer is characterized by malignant tumors arising from the epithelium covering the upper aerodigestive tract, and the majority of these epithelial malignancies are squamous cell carcinomas (SCCs) of the oral cavity (OSCCs). The aim of the current work was to identify miRNAs regulated in OSCC cancerous tissue when compared to a healthy adjacent tissue and to verify the presence of the same miRNAs in the circulation of these patients. For that serum samples and biopsies of healthy and tumor tissues were collected from five patients diagnosed with OSCC of the oral cavity, RNA was extracted from these samples and microRNAs libraries were prepared and sequenced. A total 255 miRNAs were identified in tissue and 381 different miRNAs were identified in serum samples. When comparing the miRNA expression between tumor and healthy tissue we identified 48 miRNAs (25 down- and 23 up-regulated) that were differentially expressed (FDR < 0.05). From these 48 differentially expressed miRNAs in tissue, 30 miRNAs were also found in the serum of the same patients. hsa-miR-32-5p was up-regulated in tumor compared to healthy tissue in our study, and was previously shown to be up-regulated in the serum of OSCC patients. Therefore, this suggests that miRNAs can be used as potential non-invasive biomarkers of OSCC.

Immunophenotype of neutrophils in oral squamous cell carcinoma patients.

BACKGROUND: The aim was to explore the immunophenotype of neutrophils and lymphocytes and the inflammatory mediators in patients with oral squamous cell carcinoma, comparing with controls; and to associate with clinicopathological data. METHODS: Blood was collected from 13 patients and 13 controls. The immunophenotype of neutrophils (CD66b, CD16, CD11a, arginase-1), T lymphocytes (CD4, CD8) and the intracellular cytokine production (IL-10, TNF, IFN-γ) was evaluated by flow cytometry. Plasma concentration of sVCAM-1, sTNF-RI, sTNF-RII, and IL-1ß was measured by ELISA. MPO, Lipocalin-2/NGAL, sICAM-1, and p-selectin were quantified by Luminex assay. The excised tumors were submitted to immunohistochemistry for neutrophils (CD66b) and lymphocytes (CD3, CD4, CD8). Association with clinical data was explored. P values <.05 were considered significant. RESULTS: Patients presented higher percentage of neutrophils and lower lymphocytes, resulting a higher neutrophil/lymphocyte ratio than controls. They also presented higher percentage of neutrophils expressing CD66b+ , CD66b+ Arginase-1+ , CD66b+ IL10+ , CD66b+ TNF+ , CD66b+ Arginase-1+ IL-10+ , and lower CD66b+ CD16+ CD11a+ and CD66b+ Arginase-1+ TNF+ . CD66b+ neutrophils were detected in all tumors, with a CD66b+ /CD3+ ratio of 0.40. Patients showed higher concentration of plasmatic sVCAM-1 and lower Lipocalin-2/NGAL. Patients with good outcome presented lower percentage of neutrophils, higher percentage of lymphocytes, and lower NLR than patients who died. CONCLUSION: The amount and immunophenotype of neutrophils and lymphocytes differ between patients and healthy individuals, with a pro-tumorigenic profile of neutrophils. As these cells also get within tumor microenvironment, they possibly exert systemic and local functions in cancer pathogenesis. The association of neutrophil count with outcome corroborates recent studies and this merits further investigation for applicability as a prognosticator.

Assessment of serum copper, iron and immune complexes in potentially malignant disorders and oral cancer.

Potentially malignant disorders (PMDs) of oral cavity and oral cancer remain a cause of serious concern despite intensive research and development. Diet and immunity have been identified to play a crucial role as modifying factors in these diseases. Our study intended to explore this relationship by estimating and comparing the serum levels of copper, iron and circulating immune complexes (CICs) in patients diagnosed with PMDs and oral cancer and normal healthy individuals. In this study, 40 histopathologically diagnosed cases of PMDs and oral cancer were included along with 30 healthy controls and 5 ml of venous blood was drawn using venipuncture. Serum estimation of copper, iron and CIC then followed using the colorimetric and spectrophotometric methods. The data obtained was subjected to statistical analysis using one way ANOVA and Pearson's Product-Moment Correlation Test. The mean serum copper level was measured as 138.98 ± 10.13µg/100ml in the PMD group and 141.99 ± 21.44 µg/100ml in the oral cancer as compared to 105.5 + 18.81µ/100ml in the controls. The mean serum CIC levels was highest in the oral cancer (9.65 ± 0.16OD470) followed by the PMD group (0.18 + 0.21 OD470) and least in the control group (0.048 ± 0.02OD470). Whereas, the serum levels of iron showed a significant decrease in the PMD group (110.9 ± 10.54 µg/100ml) and the oral cancer group (114.29 ± 25.83 µg/100ml) as compared with the control group (136.85 ± 14.48 µg/100ml). There was no positive correlation obtained between the three groups with respect to the chosen parameters indicating that the variables were independent of each other. It can be thus be ascertained that trace elements like copper and iron as well as humoral responses (CICs) have a close relationship with PMDs and oral cancers.

Assessment of serum copper, iron and immune complexes in potentially malignant disorders and oral cancer

Abstract Potentially malignant disorders (PMDs) of oral cavity and oral cancer remain a cause of serious concern despite intensive research and development. Diet and immunity have been identified to play a crucial role as modifying factors in these diseases. Our study intended to explore this relationship by estimating and comparing the serum levels of copper, iron and circulating immune complexes (CICs) in patients diagnosed with PMDs and oral cancer and normal healthy individuals. In this study, 40 histopathologically diagnosed cases of PMDs and oral cancer were included along with 30 healthy controls and 5 ml of venous blood was drawn using venipuncture. Serum estimation of copper, iron and CIC then followed using the colorimetric and spectrophotometric methods. The data obtained was subjected to statistical analysis using one way ANOVA and Pearson's Product-Moment Correlation Test. The mean serum copper level was measured as 138.98 ± 10.13µg/100ml in the PMD group and 141.99 ± 21.44 µg/100ml in the oral cancer as compared to 105.5 + 18.81µ/100ml in the controls. The mean serum CIC levels was highest in the oral cancer (9.65 ± 0.16OD470) followed by the PMD group (0.18 + 0.21 OD470) and least in the control group (0.048 ± 0.02OD470). Whereas, the serum levels of iron showed a significant decrease in the PMD group (110.9 ± 10.54 µg/100ml) and the oral cancer group (114.29 ± 25.83 µg/100ml) as compared with the control group (136.85 ± 14.48 µg/100ml). There was no positive correlation obtained between the three groups with respect to the chosen parameters indicating that the variables were independent of each other. It can be thus be ascertained that trace elements like copper and iron as well as humoral responses (CICs) have a close relationship with PMDs and oral cancers.

Serum big endothelin-1 as a biomarker in oral squamous cell carcinoma patients: an analytical study.

UNLABELLED: Detection of abnormally elevated levels of molecules in patients with oral cancer may be useful in early diagnosis. These markers can be included in current Histopathology grading and in TNM staging systems of Oral Squamous Cell Carcinoma (OSCC) to make it more efficient. Several pro-angiogenic molecules have been assessed for the same reason. Endothelin-1 (ET-1) is a vasoactive peptide associated with the development and spread of many solid tumors, including Squamous Cell Carcinoma (SCC), but its utility in OSCC has not been confirmed. OBJECTIVE: This study aims to evaluate the role of the serum big ET-1 as a biomarker of OSCC, by correlating it with the clinical staging and the histopathological grading. MATERIAL AND METHODS: Serum levels of big ET-1 measured by the sandwich Enzyme-Linked Immunosorbent Assay (ELISA) in 40 OSCC cases were compared with the levels from the control group using independent t-test. Clinical stages and histopathological grades of OSCC cases were compared in relation to their mean levels of serum big ET-1, one using the Analysis of Variance (ANOVA) test and the other the independent t-test, respectively. The significance of the mean difference between the groups was evaluated by Tukey's multiple comparison test. All statistical analyses were performed on GraphPad statistical software version 5.0. RESULTS: By comparing the mean of the big ET-1 concentrations of cases and controls, the independent t-test revealed significant higher big ET-1 concentration of OSCC cases when compared to controls (p<0.0001). Tukey's multiple comparison test also revealed statistically significant difference among all OSCC stages in relation to the mean levels of serum big ET-1. However, the mean of the big ET-1 concentrations of cases of grade I and of grade II did not differ statistically (p=0.729). CONCLUSION: Serum big ET-1 levels may be useful as a diagnostic tool in OSCC and as an adjunct to OSCC staging. However, its use as a prognostic marker warrants larger prospective studies.

Serum big endothelin-1 as a biomarker in oral squamous cell carcinoma patients: an analytical study

Detection of abnormally elevated levels of molecules in patients with oral cancer may be useful in early diagnosis. These markers can be included in current Histopathology grading and in TNM staging systems of Oral Squamous Cell Carcinoma (OSCC) to make it more efficient. Several pro-angiogenic molecules have been assessed for the same reason. Endothelin-1 (ET-1) is a vasoactive peptide associated with the development and spread of many solid tumors, including Squamous Cell Carcinoma (SCC), but its utility in OSCC has not been confirmed.Objective This study aims to evaluate the role of the serum big ET-1 as a biomarker of OSCC, by correlating it with the clinical staging and the histopathological grading.Material and Methods Serum levels of big ET-1 measured by the sandwich Enzyme-Linked Immunosorbent Assay (ELISA) in 40 OSCC cases were compared with the levels from the control group using independent t-test. Clinical stages and histopathological grades of OSCC cases were compared in relation to their mean levels of serum big ET-1, one using the Analysis of Variance (ANOVA) test and the other the independent t-test, respectively. The significance of the mean difference between the groups was evaluated by Tukey’s multiple comparison test. All statistical analyses were performed on GraphPad statistical software version 5.0.Results By comparing the mean of the big ET-1 concentrations of cases and controls, the independent t-test revealed significant higher big ET-1 concentration of OSCC cases when compared to controls (p<0.0001). Tukey’s multiple comparison test also revealed statistically significant difference among all OSCC stages in relation to the mean levels of serum big ET-1. However, the mean of the big ET-1 concentrations of cases of grade I and of grade II did not differ statistically (p=0.729).Conclusion Serum big ET-1 levels may be useful as a diagnostic tool in OSCC and as an adjunct to OSCC staging. However, its use as a prognostic marker warrants larger prospective studies.

Small RNAs in metastatic and non-metastatic oral squamous cell carcinoma.

BACKGROUND: Small non-coding regulatory RNAs control cellular functions at the transcriptional and post-transcriptional levels. Oral squamous cell carcinoma is among the leading cancers in the world and the presence of cervical lymph node metastases is currently its strongest prognostic factor. In this work we aimed at finding small RNAs expressed in oral squamous cell carcinoma that could be associated with the presence of lymph node metastasis. METHODS: Small RNA libraries from metastatic and non-metastatic oral squamous cell carcinomas were sequenced for the identification and quantification of known small RNAs. Selected markers were validated in plasma samples. Additionally, we used in silico analysis to investigate possible new molecules, not previously described, involved in the metastatic process. RESULTS: Global expression patterns were not associated with cervical metastases. MiR-21, miR-203 and miR-205 were highly expressed throughout samples, in agreement with their role in epithelial cell biology, but disagreeing with studies correlating these molecules with cancer invasion. Eighteen microRNAs, but no other small RNA class, varied consistently between metastatic and non-metastatic samples. Nine of these microRNAs had been previously detected in human plasma, eight of which presented consistent results between tissue and plasma samples. MiR-31 and miR-130b, known to inhibit several steps in the metastatic process, were over-expressed in non-metastatic samples and the expression of miR-130b was confirmed in plasma of patients showing no metastasis. MiR-181 and miR-296 were detected in metastatic tumors and the expression of miR-296 was confirmed in plasma of patients presenting metastasis. A novel microRNA-like molecule was also associated with non-metastatic samples, potentially targeting cell-signaling mechanisms. CONCLUSIONS: We corroborate literature data on the role of small RNAs in cancer metastasis and suggest the detection of microRNAs as a tool that may assist in the evaluation of oral squamous cell carcinoma metastatic potential.

Gene expression patterns through oral squamous cell carcinoma development: PD-L1 expression in primary tumor and circulating tumor cells.

Oral squamous cell carcinoma (OSCC) is the most common tumor of the oral cavity and has been associated with poor prognosis. Scarce prognostic markers are available for guiding treatment and/or sub-classifying patients. This study aims to identify biomarkers by searching for genes whose expression is increased or decreased during tumor progression (through T1 to T4 stages). Thirty-six samples from all tumor size stages (from T1 to T4) were analyzed using cDNA microarrays. Selected targets were analyzed by immunohistochemistry and in circulating tumor cells by immunofluorescence and Nanostring. Correlation was shown between PD-L1 and tumor size and lymph node metastasis, HOXB9 and tumor size, BLNK and perineural invasion, and between ZNF813 and perineural invasion. PD-L1 positivity was an independent prognostic factor in this cohort (p = 0.044, HH = 0.426). In CTCs from patients with locally advanced OSCC, we found a strong cytoplasmatic expression of PD-L1. PD-L1 is a ligand of PD-1 and is believed to limit T cell activity in inflammatory responses and limit autoimmune diseases. We demonstrated an important role for PD-L1 in primary tumors according to tumor size, and in disease specific survival. Therefore, we could further determine individuals with PD-L1+ CTCs, and possibly follow treatment using CTCs.

El sistema lewis hoy. nuevos enfoques / The lewis system today. new approaches

La expresión de antígenos (Ags) Lewis depende de alelos heredados en loci independientes, el gen Secretor (SE) que codifica la fucosiltransferasa 2 (FUT2) y el gen Lewis (LE) que codifica la fucosiltransferasa 3 (FUT3). El gen Se codifica una glicosiltransferasa que adiciona una fucosa en la cadena precursora de tipo 1 formando el Ag H en secreciones y fluidos. Como los azúcares inmunodominantes del Ag A y B pueden ser agregados a la cadena H de tipo 1, la FUT2 también controla la expresión de Ag A y B en las secreciones. El gen se es un alelo no funcional. El gen Le codifica una transferasa diferente que adiciona una fucosa en el 2do carbono en el precursor de tipo 1. El alelo le no es funcional. Las FUT2 y FUT3 interactúan para la formación de Ags Lewis en secreciones y fluídos. Los Ags Lewis en los eritrocitos no son en realidad parte integral de la membrana, están adsorbidos sobre la superficie en forma pasiva a partir del plasma. Están ampliamente distribuidos en tejidos humanos, eritrocitos, endotelio, riñón, tracto genitourinario, epitelio gastrointestinal y son receptores para algunos patógenos. Los anticuerpos (Acs) anti-Lewis en general no son clínicamente significativos, aunque se han publicado algunos casos de reacciones transfusionales hemolíticas, enfermedad hemolítica fetoneonatal y rechazo de transplante renal. Este trabajo es una revisión sobre los Ags del Sistema Lewis enfocada hacia sus diferentes funciones biológicas y su importancia en campos variados fuera del Banco de Sangre y la Inmunohematología tradicional.

The expression of Lewis blood group antigens depends on the alleles inherited at independent loci, FUT2 Secretor gene (SE) and FUT3 Lewis gene (LE). The Se and Le alleles encode separate fucosyltransferases that interact to form Lewis antigens in secretions and fluids. The Lewis antigens on red blood cells are not integral to the membrane but are passively adsorbed from the plasma. The allele Se encodes a transferase that adds fucose to type 1 precursor chains in secretions and fluids to form type 1 H antigen. Because A and B terminal sugars may be added to type 1 H chains, FUT2 also controls A and B expression in secretions. The FUT2 allele se gen is a nonfunctional allele. The FUT3 allele Le encodes a transferase that adds a fucose in other position in type 1 H precursor. The FUT3 allele le gen is a nonfunctional allele. The Le antigens are widely distributed in human tissues and fluids and are receptors for some pathogenic organisms. Lewis antibodies are rare and clinically no significant, although there are some reports of hemolytic transfusion reactions, hemolytic disease of the newborn and renal transplant rejection. This review focuses on different biological functions of Lewis antigens and their importance in some fields other than Blood Banks and traditional.

Expression of the gene encoding secretor type galactoside 2 α fucosyltransferase (FUT2) and ABH antigens in patients with oral lesions.

OBJECTIVE: The aim of this work was to evaluate the expression of FUT2 gene in saliva and histo ABH antigens of patients with oral lesions. STUDY DESIGN: In total 178 subjects were examined, half of whom suffered from oral pre-cancerous and cancerous lesions, while the other half were the healthy control group We analyzed the FUT 2 polymorphism by ASO-PCR (allele specific oligonucleotid - polymerase chain reaction) with specific primers for G428 allele and the wild type allele of FUT2 gene. To reveal A, B and H antigens in tissue sections of the patients (n= 89) we used a modified specific red cell adherence technique. RESULTS: We found a high intensity of oral disease in the non-secretor group (OR = 2.43). A total of 58% of the patients with oral pre-cancerous and cancerous lesions was non secretors (se_/_), in contrast with the healthy population (21.5%). A strongly positive reaction was defined as a sheet of indicator erythrocytes adhered to the epithelial cells. In 31 of the 54 samples analyzed the test showed slightly positive results on atypical areas, and there was a complete antigen deletion in areas affected by neoplasia. Nineteen samples showed a total absence of ABH antigens in both histologically normal and pathological areas. Blood group antigens were expressed at a high level in benign and highly differentiated malignant tumors. In poorly differentiated malignant tumors, they were mostly absent. CONCLUSION: Considering these results we suggest the use of this method to monitor probable preneoplastic lesions in risk population, especially in those with no secretor status (absence of FUT2 gene).

Serum albumin in poorly differentiated, oral squamous cell carcinoma: a possible diagnostic marker of significance / Albumina sérica em carcinomas de células orais escamosas e pobremente diferenciadas: um possível marcador de diagnóstico de significância

Objectives: Serum albumin is considered to be the most potent and abundant extra-cellular anti-oxidant thatmight have a protective role in the ongoing process of transition of the various oral pre-cancerous lesions andconditions into frank malignant degenerations. The aim of this study was to check the reliability of serum albuminas one of the diagnostic anti-oxidant parameter. Materials and methods: The study consisted of seraanalysis of albumin in the age and sex matched normal healthy adults and patients with histologically proven,poorly differentiated oral squamous cell carcinoma. The results were analyzed using Student’s t-test andwere averaged as mean ± standard deviation. In above test, p-values less than 0.05 were taken to be statisticallysignificant. The normality of data was checked before the statistical analysis was performed. Results:The study revealed variations in sera levels of albumin to be statistically significant with the mean level ofsera albumin to be 4.956 ± 1.0579 in controls as against 3.6933 ± 1.2177 in patients with histologically proven,poorly differentiated, oral squamous cell carcinoma. Conclusions: The results of the study emphasize theneed for more studies with larger sample sizes to be conducted before a conclusive role could be drawn infavor of sera levels of albumin as diagnostic markers of significance in oral squamous cell carcinoma.

Objetivos: A albumina sérica é considerada o antioxidante extracelular mais poderoso e mais abundante, que pode exercer um papel protetor no processo de transição das várias lesões e circunstâncias pré-cancerígenasorais em degenerações malignas. O objetivo deste estudo foi verificar a confiabilidade da albumina sérica como um dos parâmetros de diagnóstico antioxidante. Materiais e métodos: A análise consistiu no estudo da albumina sérica em pacientes adultos saudáveis normais, separados por idade e sexo, e em pacientes com carcinoma oral de células escamosas pobremente diferenciadas com evidência histológica comprovada. Os resultados foram analisados usando o teste t de Student, e as médias foram calculadas com ± desvio-padrão. No teste citado, os valores de p menores que 0,05 foram considerados estatisticamente significantes. A normalidade dos dados foi verificada antes da realização da análise estatística. Resultados: O estudo revelou diferenças estatisticamente significantes nos níveis de albumina sérica com o nível médio de 4.956 ± 1.0579 nos controle sem contraste com 3.6933 ± 1.2177 nos pacientes com carcinoma oral de células escamosas pobremente diferenciadas.Conclusões: Os resultados do estudo enfatizaram a necessidade de mais estudos com tamanhos de amostra maiores antes que um papel conclusivo possa ser atribuído, em favor dos níveis de albumina sérica,como marcador diagnóstico para o carcinoma oral de células escamosas.

The role of eosinophils and eosinophil cationic protein in oral cancer: a review.

Eosinophils are multifunctional leukocytes implicated in the pathogenesis of numerous inflammatory processes, such as allergies and parasitic infections. Increased number of these cells has been described in many human cancers, including oral squamous cell carcinoma, and its presence related to favourable as well as unfavourable prognosis. Although the exact role of eosinophils in tumours is not yet defined, the anti-tumour activity of these rare granulocytes has been associated with the release of their cytotoxic proteins, especially the eosinophil cationic protein, a single polypeptide chain with a molecular mass ranging from 15 to 22kDa encoded by the RNSE3 gene located on chromosome 14q11.2. Functional studies have implicated ECP in numerous processes, such as tissue remodelling in allergic inflammation; however its most striking function is the cytotoxic activity. The aim of this review is to summarise the role and functions of eosinophils and their granule-derived products in oral malignant tumours and the clinical value of the tumour-associated tissue eosinophilia for patients' prognosis.

Patients with oral squamous cell carcinoma are characterized by increased frequency of suppressive regulatory T cells in the blood and tumor microenvironment.

Oral squamous cell carcinoma (OSCC) is a cancerous lesion with high incidence worldwide. The immunoregulatory events leading to OSCC persistence remain to be elucidated. Our hypothesis is that regulatory T cells (Tregs) are important to obstruct antitumor immune responses in patients with OSCC. In the present study, we investigated the frequency, phenotype, and activity of Tregs from blood and lesions of patients with OSCC. Our data showed that >80% of CD4(+)CD25(+) T cells isolated from PBMC and tumor sites express FoxP3. Also, these cells express surface Treg markers, such as GITR, CD45RO, CD69, LAP, CTLA-4, CCR4, and IL-10. Purified CD4(+)CD25(+) T cells exhibited stronger suppressive activity inhibiting allogeneic T-cell proliferation and IFN-gamma production when compared with CD4(+)CD25(+) T cells isolated from healthy individuals. Interestingly, approximately 25% of CD4(+)CD25(-) T cells of PBMC from patients also expressed FoxP3 and, although these cells weakly suppress allogeneic T cells proliferative response, they inhibited IFN-gamma and induced IL-10 and TGF-beta secretion in these co-cultures. Thus, our data show that Treg cells are present in OSCC lesions and PBMC, and these cells appear to suppress immune responses both systemically and in the tumor microenvironment.

Expression of annexin A1 mRNA in peripheral blood from oral squamous cell carcinoma patients.

Several studies have been suggesting annexin A1 protein as an active player in tumorigenesis of many organs. Nevertheless, its tumor biomarker role has been mainly studied in tissues by immunohistochemistry or cell culture. Hence, in this investigation, the peripheral blood from 27 oral squamous cell carcinoma (OSCC) patients and 25 negative control individuals were examined by quantitative real-time PCR. Down-regulated ANXA1 expression at mRNA level was observed in OSCC samples (p=0.026). Significantly diminished mRNA levels correlated to age, sex and the anatomical site of the tumor lesion were observed. Moreover, the ROC curve analysis revealed the performance of ANXA1 expression as a suitable biomarker for patients with oral cavity cancer, especially those with 60years of age or older and/or women. For the first time, ANXA1 mRNA is revealed as blood-based biomarker, and its adoption for complementary non-invasive diagnosis of OSCC is suggested. These results suggest that, beyond the anti-inflammatory function, annexin A1 may also play a tumor suppressor role in peripheral blood cells, such as leukocytes.

Expression of vascular endothelial growth factor-C does not predict occult lymph-node metastasis in early oral squamous cell carcinoma.

Strong vascular endothelial growth factor-C (VEGF-C) expression has been correlated to occurrence of lymph-node metastases in patients with oral squamous cell carcinoma (OSCC). The incidence of occult lymph-node metastasis remains a decisive factor in the prognosis of patients with early OSCC. The aim of this study was to evaluate VEGF-C expression as a predictor of occult lymph-node metastasis in OSCC. Eighty-seven patients with primary OSCC arising in the tongue or floor of mouth, clinically T1N0M0 or T2N0M0, with (pN+) and without (pN0) occult lymph-node metastases were analyzed for VEGF-C expression by malignant cells. Occult lymph-node metastases (pN+) were detected in 22% of the 64 patients who were submitted to elective neck dissection. No statistically significant difference was found between OSCC with and without occult lymph-node metastasis in regard to VEGF-C immunoexpression by malignant cells and clinicopathologic features. Independently of VEGF-C expression, lymph-node metastasis (pN+) was the most significant prognostic factor for overall survival of patients with OSCC (p=0.030). These findings indicate that isolated VEGF-C expression by malignant cells is not of predictive value for occult lymph-node metastasis in the early stages of OSCC.

Expresión antigénica ABH y carácter secretor en pacientes con lesiones orales precancerosas y cancerosas / Secretor status and ABH antigens expression in patients with oral lesion

Los antígenos ABH, productos de la interacción de dos sistemas genéticos Hh y ABO, están sujetos a leyes de herencia y pueden estar localizados no sólo en los eritrocitos sino también en la mayoría de las células humanas. El objetivo del este trabajo fue investigar la relación entre el carácter secretor de pacientes con lesiones orales pre-malignas y malignas y la expresión antigénica ABH en cortes histológicos de dichas lesiones. Se trabajó con muestras incluídas en tacos de parafina de pacientes con lesiones orales. Los pacientes fueron clasificados en 2 grupos a) lesiones pre-malignas y malignas y b) lesiones benignas. Se investigaron los antígenos ABH por la técnica de inmunoadherencia específica modificada. Se utilizó la adherencia al tejido vascular como control positivo y al tejido adiposo como control negativo. Los resultados fueron semicuantificados desde adherencia fuertemente positiva a negativa. El carácter secretor fue determinado por la técnica de inhibición de la hemaglutinación. En 21 de las 34 muestras se observó una débil expresión antigénica en áreas atípicas, y deleción total en las áreas histológicamente afectadas por neoplasia. En 8 muestras hubo pérdida total de los antígenos ABH tanto en áreas normales como patológicas, estos pacientes presentaron un mayor grado de malignidad y metástasis que aquellos que conservaron la antigenicidad. Los pacientes con lesiones orales pre-malignas y malignas presentaron un incremento del carácter no secretor (52,3 por ciento) respecto de la población control (19,5 por ciento) y de aquellos pacientes con lesiones orales benignas (15.4 por ciento). Se observó una importante asociación entre pacientes no secretores y deleción de los antigenos ABH en muestras de lesiones orales. Además, hemos encontrado, en el grupo no secretor, una mayor malignidad de las lesiones orales como así también una mayor presentación de displasia epitelial. El estudio del carácter secretor en los pacientes con lesiones orales...(AU)

Expresión antigénica ABH y carácter secretor en pacientes con lesiones orales precancerosas y cancerosas / Secretor status and ABH antigens expression in patients with oral lesion

Los antígenos ABH, productos de la interacción de dos sistemas genéticos Hh y ABO, están sujetos a leyes de herencia y pueden estar localizados no sólo en los eritrocitos sino también en la mayoría de las células humanas. El objetivo del este trabajo fue investigar la relación entre el carácter secretor de pacientes con lesiones orales pre-malignas y malignas y la expresión antigénica ABH en cortes histológicos de dichas lesiones. Se trabajó con muestras incluídas en tacos de parafina de pacientes con lesiones orales. Los pacientes fueron clasificados en 2 grupos a) lesiones pre-malignas y malignas y b) lesiones benignas. Se investigaron los antígenos ABH por la técnica de inmunoadherencia específica modificada. Se utilizó la adherencia al tejido vascular como control positivo y al tejido adiposo como control negativo. Los resultados fueron semicuantificados desde adherencia fuertemente positiva a negativa. El carácter secretor fue determinado por la técnica de inhibición de la hemaglutinación. En 21 de las 34 muestras se observó una débil expresión antigénica en áreas atípicas, y deleción total en las áreas histológicamente afectadas por neoplasia. En 8 muestras hubo pérdida total de los antígenos ABH tanto en áreas normales como patológicas, estos pacientes presentaron un mayor grado de malignidad y metástasis que aquellos que conservaron la antigenicidad. Los pacientes con lesiones orales pre-malignas y malignas presentaron un incremento del carácter no secretor (52,3 por ciento) respecto de la población control (19,5 por ciento) y de aquellos pacientes con lesiones orales benignas (15.4 por ciento). Se observó una importante asociación entre pacientes no secretores y deleción de los antigenos ABH en muestras de lesiones orales. Además, hemos encontrado, en el grupo no secretor, una mayor malignidad de las lesiones orales como así también una mayor presentación de displasia epitelial. El estudio del carácter secretor en los pacientes con lesiones orales...

The hamster cheek pouch as a model of oral cancer for boron neutron capture therapy studies: selective delivery of boron by boronophenylalanine.

Herein we propose and validate the hamster cheek pouch model of oral cancer for boron neutron capture therapy (BNCT) studies. This model serves to explore new applications of the technique, study the biology and radiobiology of BNCT, and assess the uptake of boron compounds and response of tumor, precancerous tissue, and clinically relevant normal tissues. These issues are central to evaluating and improving the therapeutic gain of BNCT. The success of BNCT is dependent on the absolute amount of boron in the tumor, and the tumor:blood and tumor:normal tissue boron concentration ratios. Within this context, biodistribution studies are pivotal. Tumors were induced in the hamsters with a carcinogenesis protocol that uses dimethyl-1,2-benzanthracene and mimics spontaneous tumor development in human oral mucosa. The animals were then used for biodistribution and pharmacokinetic studies of boronophenylalanine (BPA). Blood, tumor, precancerous pouch tissue surrounding tumor, normal pouch tissue, tongue, skin, cheek mucosa, palate mucosa, liver, and spleen, were sampled at 0-12 h after administration of 300 mg BPA/kg. The data reveal selective uptake of BPA by tumor tissue and, to a lesser degree, by precancerous tissue. Mean tumor boron concentration was 36.9 +/- 17.5 ppm at 3.5 h and the mean boron ratios were 2.4:1 for tumor:normal pouch tissue and 3.2:1 for tumor:blood. Higher doses of BPA (600 and 1200 mg BPA/kg) increased tumor uptake. Potentially therapeutic absolute boron concentrations, and tumor:normal tissue and tumor:blood ratios can be achieved in the hamster oral cancer model using BPA as the delivery agent.